Glioblastoma (GBM) is one of the most aggressive and lethal types of human cancer. Standard treatment involves surgery followed by combined chemoradiation. However, the molecular mechanisms underlying innate tumor radioresistance remain unclear. Here, researchers found that Smoothened (Smo) expression is significantly upregulated in recurrent GBM cell lines and tumor tissues after radiotherapy. Increased Smo expression is associated with a poor prognosis in GBM patients treated with radiotherapy. Smo conferred radioresistance in both GBM cells and human tumor xenografts. The underlying mechanism of these effects involves impaired repair of radiotherapy-induced DNA damage. Importantly, Smo's effect on radioresistance is mediated by claspin polyubiquitination and proteasomal degradation, thereby regulating ATR-Chk1 signaling. Furthermore, Smo reduces claspin polyubiquitination and proteasomal degradation by promoting USP3 transcription. Researchers also demonstrated that the Smo inhibitor GDC-0449 induces radiosensitivity in GBM. These findings reveal the underlying mechanism of GBM radioresistance and suggest a potential therapeutic target for radiation resistance in GBM.
To investigate whether Smo is associated with radioresistance, the researchers generated Smo-overexpressing primary GBM cells (survival rate less than 45% after 2 Gy irradiation). As expected, Smo was overexpressed in both primary GBM cell lines (Figure 1A). They then investigated the effect of Smo overexpression on the survival of primary GBM cells after irradiation (IR). As shown in Figure 1B, Smo overexpression enhanced the resistance of primary GBM cells to IR. Next, they assessed the apoptosis rate of Smo-overexpressing and control cells after IR. The results showed that compared with control cells, Smo-overexpressing cells had significantly fewer apoptotic cells (Annexin V-PI/FITC or TUNEL staining) and significantly decreased levels of cleaved caspase-3 (Figures 1C-E). These results indicate that Smo overexpression induces resistance to IR in primary GBM cells and protects them from IR-induced apoptosis.
Figure 1. Smo overexpression confers radiation resistance. (Tu Y, et al., 2020)